Macroscopic cartilage formation with embryonic stem-cell-derived mesodermal progenitor cells

Abstract

The totipotent embryonic stem cell generates various mesodermal cells when stimulated with BMP4. Among the resulting cells, those expressing flk-1 and/or PDGFRα displayed chondrogenic activity in the presence of TGFβ3 and expressed cartilage-specific genes in 7 to 16 day pellet cultures. Depositions of cartilage matrix and type II collagen were detected by day 14. TGFβ-stimulated chondrogenesis was synergistically enhanced by PDGF-BB, resulting in a larger cartilage particle filled with a cartilaginous area containing type II collagen, with a surface cell layer expressing type I collagen. In contrast, noggin inhibited both the TGFβ-and TGFβ+PDGF-stimulated cartilage formation, suggesting that a BMPdependent pathway is involved. In fact, replacement of TGFβ3 with BMP4 on days 10 to 12 markedly elevated the cartilage matrix deposition during the following 7 to 8 days. Moreover, culture with TGFβ3 and PDGF-BB, followed by the incubation with BMP4 alone, resulted in a cartilage particle lacking type I collagen in the matrix and the surface layer, which suggests hyaline cartilage formation. Furthermore, such hyaline cartilage particles were mineralized. These studies indicate that the PDGFRα + and/or flk-1 + cells derived from embryonic stem cells possess the full developmental potential toward chondrocytes, in common with embryonic mesenchymal cells.

Cartilage tissue engineering using differentiated and purified induced pluripotent stem cells

“…A nonuniform cell population not only limits the effectiveness of the therapy or model, but also increases the risk for undifferentiated cells contained in the population to contribute to teratoma formation (18). Previous work with mouse and human embryonic stem cells has demonstrated that chondrogenesis of pluripotent cells can be initiated by exposure to growth factors such as the bone morphogenetic proteins (BMPs) (19,20) and transforming growth factor-beta (TGF-β) (21,22). Chondrogenesis can be further enhanced by recapitulating features of mesenchymal condensation using 3D culture systems such as embryoid bodies (23), micromasses (24,25), pellets (26), or scaffolds (27).…”

Tissue engineering of blood vessels with endothelial cells differentiated from mouse embryonic stem cells

“…It has been reported that ES cells can differentiate into many kinds of committed cells by the induction with growth factors, cytokines or conditioned culture medium, and by gene manipulation [8,18,19], etc. Therefore, ES cells are expected to become a new source of seed cells for tissue engineering.…”

Skeletal tissue engineering using embryonic stem cells

“…However, in the in vitro samples we sometimes observed chondrocytes in large lacunae surrounded by a thin layer of cartilage ECM, a morphology observed in hypertrophic cartilage. It has been reported by other groups that mouse and human ESCs express hypertrophic markers and even some osteogenic markers in vitro [88,101,155]. Therefore, we would have to further investigate if the ESC-derived cartilage already displayed signs of hypertrophy before implantation.…”